A tool kit for molecular genetics of Kluyveromyces lactis comprising a congenic strain series and a set of versatile vectors

Autor(en): Heinisch, Juergen J.
Buchwald, Ulf
Gottschlich, Andre
Heppeler, Nele
Rodicio, Rosaura
Stichwörter: Biotechnology & Applied Microbiology; bud-site selection; BUDDING YEAST; carbohydrate metabolism; CASSETTES; Cre-recombinase; EVOLUTION; GENES; GLUCOSE REPRESSION; homologous recombination; ISOCITRATE LYASE; Kluyveromyces lactis CBS2359; MARKER; Microbiology; Mycology; PLASMIDS
Erscheinungsdatum: 2010
Herausgeber: OXFORD UNIV PRESS
Journal: FEMS YEAST RESEARCH
Volumen: 10
Ausgabe: 3
Startseite: 333
Seitenende: 342
Zusammenfassung: 
A set of different marker deletions starting with a ura3 derivative of the Kluyveromyces lactis type strain CBS2359 was constructed. After a first cross to obtain a strain with the opposite mating type that also carried a leu2 allele, continuous back-crosses were used to obtain a congenic strain series with different marker combinations, including deletions in KlHIS3, KlADE2 and KlLAC4. Enzymes involved in carbohydrate metabolism were shown to behave very similarly to the original type strain and other K. lactis strains investigated previously. Moreover, a vector series of Saccharomyces cerevisiae genes flanked by loxP sites was constructed to be used as heterologous deletion cassettes in K. lactis, together with two plasmids for expression of Cre-recombinase for marker regeneration. To increase the frequency of homologous recombination, the Klku80 deletion was also introduced into the congenic strain series. A PCR-based method for determination of mating type is provided.
ISSN: 15671356
DOI: 10.1111/j.1567-1364.2009.00604.x

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